DISSERTATION - SYNOPSIS
DR. LAKSHMI ALVA .M
POSTGRADUATE STUDENT
DEPARTMENT OF ORAL AND MAXILLOFACIAL PATHOLOGY
BATCH 2013-2014
A.J.INSTITUTE OF DENTAL SCIENCES, KUNTIKANA,
MANGALORE.
Rajiv Gandhi University of Health Sciences, Karnataka,
Bangalore
ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1.
Name of the Candidate
And Address
(in block letters)
DR. LAKSHMI ALVA .M
POST GRADUATE STUDENT.
DEPARTMENT OF ORAL AND
MAXILLOFACIAL PATHOLOGY.
A.J.INSTITUTE OF DENTAL SCIENCES.
N.H 66, KUNTIKANA,
MANGALORE-575004.
2.
Name of the institution
A.J.INSTITUTE OF DENTAL SCIENCES.
N.H 66, KUNTIKANA,
MANGALORE-575004.
3.
Course of study and subject
MASTER OF DENTAL SURGERY,
ORAL AND MAXILLO FACIAL PATHOLOGY
AND MICROBIOLOGY
4.
Date of admission to course
5TH JULY 2013.
5.
Title of the topic:
“QUANTITATIVE ESTIMATION OF SERUM
PHOSPHODIESTERASE & PHOSPHOHEXOSE
ISOMERASE IN ORAL SQUAMOUS CELL
CARCINOMA”
6.
Brief resume of the intended work:
6.1 Need for the study:
Oral cancer is the eighth most common cancer worldwide. Cancer affects
approximately ten million people every year. Prevalence being high among men. In
India, the age-standardized incidence rate of oral cancer is 12.6 per 1 lakh population.
Cancer of the oral cavity includes lip, tongue, gingival, oral mucosa, oropharynx and
hypo pharynx. The clinical and histological features may help in predicting the diagnosis
of carcinomas but cannot always accurately predict their prognosis. Therefore
identification of molecular markers (or biomarkers) which can predict disease
progression is necessary for better management. Commonly enzymes are used as an aid
for diagnosis and prognosis1.
The enzymes are biological catalysts produced by the living cells. They play a
vital role in the metabolic reaction of cells. In pathological conditions there is usually
some derangement in the metabolic process which is reflected by a change in enzymatic
patterns4.
The phosphodiesterases (PDE) enzymes are responsible for the hydrolysis of the
second messengers with the fundamental role in the transduction of the intercellular
signals. Variations in PDE activity have been correlated to different pathological
mechanisms, such as cellular differentiation, apoptosis and tumor invasivity. PDEs are
also known to play a role in tumor growth by influencing angiogenesis1.
The head and neck region consist of a number of cavities with plenty of hidden
areas where a cancer may continue to grow till it is too late for curative treatment. The
need persists for some clinically easy and simple test which can suggest cancer quite
early even before the patients become symptomatic, if performed during routine physical
check up3.
Phosphohexose isomerase (PHI) is one of the key enzymes in the glycolytic
pathology. PHI catalysis the reversible conversion of glucose-6-phosphate to fructose-6phosphate.Normal serum levels of intracellular enzymes are produced due to routine
destruction of cells. Malignancy may be the cause of accelerated cell death resulting in
raised serum PHI levels.
Estimation of serum PHI levels have significant role in
diagnosis of cancer, early detection of residual growth, recurrent growth and
secondaries3, 4.
The diagnosis and prognosis of OSCC can be established by means of biopsy,
which is an invasive, time-consuming procedure and causes psychological trauma to
some patients. Thus the need of the hour is that, the performing tests must be simple,
less invasive, less time consuming, easy for interpretation, economical and yet quite
confirmatory for its diagnosis and prognosis.
6.2 Review of literature:
 Prabhu K, et al (2011) in his study found that pretreatment phosphodiesterase
levels were significantly elevated in oral cancer patients as compared with
healthy controls and also there was a significant increase in phosphodiesterase
levels with advancing stage in oral cancer patients1.
 Narang APS, et al (2001) in his study found that there was significantly increase
in level of serum Lactate dehydrogenase, Phosphohexose isomerase and
Carcinoembryonic antigen levels among OSCC patients and control group2.
 Verma PC, et al (2001) in his study found that the value of PHI levels for the
patients of head and neck cancer was significantly higher as compared to control
group3.
 Bhatia PL, et al (1977) carried out enzyme assay in patients suffering from head
and neck cancer, he found that enzyme levels were altered in all the cancer
patients, maximally in oral cancer with cervical metastases4.

Robert L, et al (1977) studied cAMP-PDE assay in lymphocyte from normal
controls-patients with benign conditions in various age groups.In lymphocytes
from untreated patients with stage 3 and stage 4 squamous cell carcinomas of the
head and neck, the specific activity or affinity of cAMP-PDE were higher than
the controls5.
 Taku Murata, et al (2009) studied PDE5 activity and PDE5A mRNA
expression in MAA cells. Effects of PDE5 inhibitors on cell growth were
determined by 3-5-2-2H-tetrazolium, inner salt (MTS) assay6.
 Markus Drees, et al (1993) evaluated the role of 3',5'-cyclic nucleotide
phosphodiesterase (PDE) in tumor cells as potential target for tumor growth
inhibition. The results suggested a correlation between PDE-IV inhibition and
growth inhibition. PDE-IV thus appears to be a potential new target for
antiproliferative treatment7.

Stuart J, et al (2004) assessed the role of phosphodiesterases (PDE) in
endothelial cell migration. It was found that selective inhibition of PDE2
inhibited migration of all human vascular endothelial cells. Inhibition of PDE4
activity decreased migration, and in conjunction with forskolin, increased cAMP
in vascular endothelial cells. PDE3 inhibition potentiated forskolin-induced
increases in cAMP and inhibited migration in vascular endothelial cells derived
from aorta and umbilical vein. PDE2, PDE3 and PDE4 inhibitors limited vascular
endothelial cell migration8.
6.3 Objectives of the study:
1. Quantitative estimation of serum Phosphodiesterase in OSCC patients.
2. Quantitative estimation of serum Phosphohexose isomerase in OSCC patients.
3. To compare serum Phosphodiesterase and serum Phosphohexose isomerase levels
of healthy adults with OSCC patients.
7
Material and methods:
7.1 Source of data:
Patients consulting the Department of Oral Medicine and
Radiology, A.J.INSTITUTE OF DENTAL SCIENCES MANGALORE. The study
would be under taken in the Department of Oral and Maxillofacial Pathology,
A.J.INSTITUTE OF DENTAL SCIENCES MANGALORE.
Inclusion criteria:
Study Group – Clinically and Histopathogically diagnosed cases of Oral squamous
Cell Carcinoma.
Control Group - Disease free individuals
Exclusion criteria:
1. Oral Squamous Cell Carcinoma patients associated with other conditions where
serum phosphodiesterase and serum phosphohexose isomerase is elevated. These
include
Liver diseases
Diabetes Mellitus
Renal disease.
Sample size:
Control Group -20 healthy individuals.
Study Group: Histopathologically confirmed cases of OSCC patients - 20 OSCC
Patients.
Armamentarium

For collecting serum samples: Disposable syringe to draw blood, Vacutainers to
collect blood, Centrifuge, Pipette will be used.
.

Deep freeze refrigerator to store the serum samples in polyethylene tubes at -20 c
until analysis.

Phosphodiesterase assay will be done using the following reagents:
 Bis PNPP (Bis-p-nitrophenyl phosphate sodium salt).
 Tris HCl Buffer.
 Paranitrophenol.
 Sodium hydroxide.

Serum Phosphodiesterase is estimated using spectrophotometer method.

Serum Phosphohexose isomerase is estimated manually by using Seliwanoff
reaction.
7.2 Methods of collection of data(including sampling procedure, if any)

Subjects consulting the Department of Oral Medicine and Radiology,
clinically & histopathologically diagnosed cases of Oral Squamous Cell
Carcinoma.

Informed consent will be taken from all the subjects included in the study.
Methodology:

Clinically and Histopathologically diagnosed cases of Oral Squamous Cell
Carcinoma will be included in the OSCC group.

Method for PDE (serum): Under all aseptic precautions about 2 ml of venous
blood will be collected from antecubital vein of study and control group into
plain sterile vacutainer with no anticoagulant. The sample is then allowed to
clot for 30 min and then centrifuged at 3000 rpm for 5 min, to separate the
serum. Immediately this serum is used for estimation of PDE by
spectrophotometer.

Method for PHI (serum): Under all aseptic precautions about 5 ml of venous
blood will be collected from antecubital vein of study and control group into
plain sterile vacutainer with no anticoagulant. The estimation of PHI in serum
is done manually using Seliwanoff reaction for ketose sugars based on
conversion of glucose-6-phosphate to fructose-6-phosphate.

The obtained mean values will be then compared between the OSCC and
control groups.
Statistical analysis:
 Mann-Whitney U Test would be used to compare mean values of PDE, PHI
and controls.
 SPSS Software Version 17.0 will be used.
7.3 Does the study require any investigations or interventions to be
Conducted on patients or other humans or animals? If so, please.
Describe briefly.
Yes, The study requires collection of Blood from study group & control group.
7.4 Has ethical clearance been obtained from your institution in case
Of 7.3?
Yes.
INVESTIGATION DESIGN:
Subjects will be interviewed and examined on the dental chair using clinical
examination tools. Suspected cases of OSCC were selected.
Histopathologically confirmed
cases
cases of OSCC were selected
Control group
Blood
withdrawal
Blood
withdrawal
Blood coagulated centrifuged and serum separated
Quantitative estimation of PDE & PHI done.
Results compared and Statistical analysis made
List of references:
1. Prabhu K, Naik D, Ray S, Vadiraja BM, Kamath A. Serum
phosphodiesterase in oral cancer. J of cancer research and therapeutics
2011;7(2):180-182.
2. Narang APS, Greval RS, Chopra H, Kalra CS. The role of two enzymes
(LDH and PHI) and a tumor marker (CEA) in the prognostic evaluation of
head and neck malignancy. Indian J of Otolaryngology and Head & Neck
Surgery 2001;53(1):1-6.
3. Verma PC, Ojha T, Yadav D, Hemani DD. Study of serum phosphohexo
isomerase (PHI) levels in the management of head and neck malignancies.
Indian J of Otolaryngology and Head & Neck Surgery 2001;53(1):1-7.
4. Bhatia PL, Singh MM, Chakravorty M. Enzymatic study in diagnosis of
head and neck cancer. Indian J of Otolaryngology and Head & Neck
Surgery 1979;31(3):76-9.
5. Banner RL, Vaughn KW, Hagey KA, Thompson WJ, Hersh EM. Cyclic
adenosine monophosphate-phoshodiesterase(c AMP-PDE) in the
lymphocytes from patients with stage 3 and 4 squamous cell carcinoma of
the head and neck. J of surgical oncol 1977;9(1):61-9.
6. Murata T, Shimizu K, Watanabe Y, Morita H, Sekida M, Tagawa T.
Expression and role of phosphodiesterase5 in human malignant melanoma
cell line. Anticancer Research 2010;30:355-8.
7. Drees M, Zimmermann R, Eisenbrand G. 3, 5-Cyclic nucleotide
phosphodiesterase in tumor cells as potential target for tumor growth
inhibition. Cancer Research 1993;53:3058-61.
8. Netherton SJ, Maurice HD. Vascular endothelial cell cyclic nucleotide
phosphodiesterases and regulated cell migration: Implications in
angiogenesis. The American Society for pharmacology and experimental
therapeutics.2005;67(1):263-72.
9.
Signature of candidate
10.
Remarks of the guide:
11.
11.1 Name & Designation DR.PREETHI B. NAYAK
of Guide:
READER
DEPARTMENT OF ORAL AND
(in block letters):
MAXILLOFACIAL PATHOLOGY.
A. J .INSTITUTE OF DENTAL SCIENCES.
MANGALORE.
SATISFACTORY
11.2 Signature:
11.3 Co-Guide (if any)
DR. DINKAR DESAI
PROFESSOR & HOD
DEPARTMENT OF ORAL AND
MAXILLOFACIAL PATHOLOGY.
A. J .INSTITUTE OF DENTAL SCIENCES.
MANGALORE.
11.4 Signature:
11.5 Head of Department
11.6 Signature
12.
12.1 Remarks of the
Chairman &Principal:
12.2 Signature
DR. DINKAR DESAI
PROFESSOR & HOD
DEPARTMENT OF ORAL AND
MAXILLOFACIAL PATHOLOGY.
A. J .INSTITUTE OF DENTAL SCIENCES.
MANGALORE.